Near infrared fluorescent probe for detecting zinc ion in cell and its synthesis method and use

Abstract

This invention is near-infrared fluorescent probe for intracellular zinc ion test. The syncetic process is as followings: A.mix cyanine dyes Cy.7 and complexing group in the weight propotion of 1:5-10, resolve the mix in N,N-dimethyl formamide, and react at 60-70deg C for 2-5 hours with the protection of inactive gas; B. Cool the production A to 10-20deg C, dry blowing with depression, resolve in aether, add water for extraction, the extraction liquor I obtained, then add acetic ester for extraction, the extraction liquor II obtained; C. Add anhydrous addex-magnesium in liquor II for desiccation, dry blowing with depression at 30-40deg C, get rid of acetic ester for the final production. The near-infrared fluorescent probe has a good cellular osmosis and little side effect. In particular, it's seligible for cellular zinc ion test.

Description

technical field [0001] The present invention relates to the fields of biological detection technology and clinical medical detection, in particular to a near-infrared fluorescent probe for detecting intracellular zinc ions; the present invention also relates to a synthesis method of the fluorescent probe; in addition, the present invention also relates to the fluorescent probe the use of. Background technique [0002] Zinc ions are the second most abundant transition metal in living organisms. A large number of zinc ions are concentrated in nerve tissue, such as the concentration in brain tissue is 0.1-0.5nM. Zinc ions are directly involved in the regulation of gene expression through metal regulatory proteins. For example, zinc, as a component of zinc finger proteins, can affect gene expression by changing the structure of chromatin or by interrupting DNA synthesis. Most of the zinc ions in organisms are tightly combined with proteins, and there are "free zinc pools" in s...

AI Technical Summary

Problems solved by technology

[0003] Many existing methods, including atomic absorption spectrometry, electron paramagnetic resonance, Electron Probe Micronalysis, etc., have been used for the detection of metal ions in cells, but the instruments used in these methods are expensive, and they cannot perform real-time and original detection of metal ions in organisms. Bit dynamic detection, and the sample pretreatment is more complicated when these methods are determined, so the application is limited

[0004] At present, existing fluorescent probes that can detect cellular zinc ions mainly include the following categories: TSQ (Frederickson, C.J.; Kasarkis, E.J.; Ringo, D.; Frederickson, R.E.J.Neurosci.Methods, 1987,20,91- 103.), Zinquin ethyl ester (Tsuda, M.Neurosci., 17, 6678, 1997) dansylaminoethyl cyclamate (Koike, T.; Watanabe, T.; Aoki, S.; Kimura, E. ; Shiro, M., J.Am.Chem.Sic., 118, 12696-12703, 1996) zinpyr (Burfrtte, S.C.; Frederickson, C.J.; Bu, W.; Lippard, S.J., J.Am.Chem.Soc. , 2003, 125, 1778-1787.) and ZnAF (Hirano, T.; Kikuchi, K.; Urano, Y.; Higuchi, T.; Nagano, T.J.Am.Chem.Soc., 2002, 124, 6555-6562 .) etc., but the disadvantage of these probes is that their excitation wavelength is in the ultraviolet region (~350nm), which will cause damage to the cells when excited, and because their emission wavelength is in the ultraviolet or visible region, the autofluorescence interference of the biological body is serious, and the pH change Sensitive, making it subject to certain restrictions

Another problem is that it is generally difficult to synthesize such probe molecules, and the structure is relatively complex, so it will be difficult for practical application.

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